KR20090008807A - Method and apparatus for accelerating growth of yeasts by applying electric field - Google Patents

Abstract

A method for promoting the growth of yeast by using the electric field is provided to reduce the time to be required for fermentation of liquor including beer and wine and bread, and increase the production efficiency of bio-ethanol. The yeast growth is promoted by applying the AC(alternating current) electric field or DC(direct current) electric field of less than 1 Hz in the intensity of 1V to 100MV/m to the yeast, wherein the frequency of the electric field is less than 1muHz. The device for promoting the growth of yeast by using the electric field comprises: a vessel(1) made of insulator; two electrodes(2) adhered to the bottom of the vessel and outer surface of the lid; and an electric field applying apparatus(3) connected to two electrodes, wherein one of two electrodes is earthed, and another electrode is connected to the anode or cathode of the electric field applying apparatus.

Description

Method and apparatus for promoting yeast growth using electric field {Method and apparatus for accelerating growth of yeasts by applying electric field}

The present invention relates to a method and apparatus for promoting yeast growth by applying an electric field to yeast by a non-contact electric field using a low frequency band electric field of 1 Hz or less, and is applicable to all yeasts used industrially, in particular, yeast In addition to increasing itself, the invention relates to methods applicable to increasing the production of all metabolites of yeast (eg ethanol, carbon dioxide).

There are a variety of industries that use yeast growth. Liquor manufacturing industry, such as beer and wine, which uses alcoholic fermentation of yeast, the food and pharmaceutical industry using yeast extract as raw material, and bioethanol manufacturing industry, which manufactures industrial ethanol using alcohol fermentation of yeast Ranging from.

Beer is produced by alcohol fermentation using yeast after undergoing saccharification process to hydrolyze the grain consisting of starch into maltose and glucose of fermentable sugars. As a raw material, barley, wheat, corn, rice, cassava or potato may be used. In the case of barley, since a large amount of amylase is formed during germination, it may be used for saccharification. After drying, store it. This dried and germinated barley is called malt. For other feedstocks, saccharification is achieved by the addition of debranching enzymes such as alpha-amylase, beta-amylase, amyloglucosidase and perlulanase. After saccharification has reached the desired stage, the mixture of grains and enzymes is stopped to stop further hydrolysis, and then the hops are added to the filtrate and fermented by inoculation with yeast.

Beer fermentation inoculates specific homo strains obtained by pre-fermentation at low temperatures for 5 to 10 days. Most of the homo strains used in beer production are Saccharomyces carlsbergensis and Saccharomyces cerevisiae. Saccharomyces cerevisiae belongs to the species (species). Saccharomyces cerevisiae yeast is used for ale type beer which is fermented at 14-23 ° C. for 5-7 days, and lager type beer which is fermented at 6-12 ° C. for 8-14 days. Saccharomyces uvarum or Saccharomyces uvarum yeast is used. Meanwhile, a variety of Torulaspora delbrueckii yeasts are used for barbarian-style wheat beer production, and natural yeast belonging to the genus Brettanomyces is used for fermentation of the Belgian beer, Lambic. . After fermentation of sugar is converted to ethanol due to fermentation, most of the yeast is removed and stored and aged for various periods of time.

Yeast is also used in winemaking to convert sugar in grape juice to alcohol. Fermentation may be carried out using natural yeast originally present in the grape skin, or fermented by inoculating certain homo strains such as Saccharomyces cerevisiae.

In baking, Saccharomyces cerevisiae yeast is also used to convert sugar in the dough to carbon dioxide. Saccharomyces exiguous, a natural yeast found in plants, fruits and grains, is also used.

Industrial ethanol can also be obtained through a fermentation process that incompletely burns biomass with yeast. Since ethanol is completely combustible and does not cause air pollution problems, it can be used as an automobile fuel instead of gasoline and also used as a synthetic raw material for industrial chemicals.

Bioethanol manufacturing process is a step of pretreating the substrate (cassava (cassava, sugar cane, sugar corn, molasses, such as molasses)) as a raw material and adding diluted sulfuric acid or amylase enzyme; Ethanol fermentation using yeast; And purification by distillation and / or dehydration. In the fermentation process, as shown in the following Scheme 1 or 2, the hexasaccharide or pentose sugar contained in the biomass is fermented by yeast and converted to ethanol, and in the case of hexasaccharide, the actual fermentation yield is about 90 of the theoretical yield. 0.46 g ethanol / g glucose.

C ₆ H ₁₂ O ₆ → 2C ₂ H ₅ OH + 2CO ₂

3C ₅ H ₁₀ O ₅ → 5C ₂ H ₅ OH + 5CO ₂

On the other hand, yeast extract is a natural seasoning containing a large amount of amino acids, peptides, inorganic salts, vitamins, nucleic acids, etc. and is used as an important food material in modern times when there is a great demand for natural foods (eg Vegemite, Marmite, Promite, Bovril). , Oxo, Cenovis). The yeast extract has a high safety, and the demand is increasing in that it has a complex aesthetic such as unique flavor, rich taste that does not have a chemical synthetic seasoning.

Inactivated yeast (especially Saccharomyces cerevisiae) is a good source of protein, vitamins (especially B-complex vitamins), minerals and cofactors, and is also used as a nutrient because of its low fat and salt.

A general method for preparing a yeast extract is a method of self-extinguishing, in which salt is added to a yeast suspension to shrink cells, and when the self-extinguishing occurs due to intrinsic enzymes in yeast cells, the reaction is completed by heating. It is also possible to use an enzyme digestion method in which various enzymes are properly added and externally added.

Saccharomyces boulardii yeast can also be used as probiotics (probiotic) to maintain beneficial microbial populations present in the large and small intestine.

In most of the industries described above, multiplying yeast in a large amount of time is directly related to the efficiency of the industry. For example, in the liquor manufacturing industry or the bakery industry using yeast fermentation, since fermentation proceeds at a lower temperature than the optimum temperature for yeast growth, the metabolism and growth rate of yeast are lowered, and the production efficiency is lowered. When ethanol accumulates, yeast is inhibited from growth, which reduces ethanol production efficiency. Therefore, there is a need for a method for promoting yeast growth that can solve the problem of reduced production efficiency.

On the other hand, the effect of applying the electric field to the yeast, there is a study results that the yeast is killed when applying an electric field having an intensity of 20-30kV / cm, a radio frequency of 20-60kHz (Geveke DJ, Brunkhorst C. Inactivation of Saccharomyces cerevisiae with radio frequency electric fields.JOURNAL OF BACTERIOLOGY, Feb. 1994, p.702-713). In addition, studies have shown that applying a pulsed electric field increases the efficiency of transfection of foreign DNA into yeast (High Efficiency Transformation of Intact Yeast Cells by Electric Field Pulses.Eliane Meilhoc, Jean-Michel Masson, & Justin Teissie. Nature Biotechnology 8, 223-227 (1990)). However, there is no known method of promoting the growth of yeast by using an electric field.

Japanese Patent Laid-Open Nos. 2004-159644 and 2004-161740 (Ultra biotech ltd.) And US Patent Nos. US6436695, US6391619, US6391618, US6391617, and US6440713 describe a method for activating yeast with a specific medical effect using an electric field, but there is no description of the proliferative effect of yeast.

The inventors have found that application of an electric field in the low frequency band below 1 Hz to yeast promotes the growth of yeast. Accordingly, it is an object of the present invention to provide a yeast growth promoting method and apparatus for applying an electric field to yeast in order to increase the production efficiency of yeast itself and its metabolites. It is also an object of the present invention to provide a method of using the yeast growth promoting method to shorten the time required for liquor and bread fermentation of existing beer, wine and the like, and to increase the efficiency of bioethanol production.

In order to solve the above problems, the present invention provides a yeast growth promoting method comprising the step of applying the electric field so that the alternating current or direct current electric field of less than 1 Hz to the yeast.

In the yeast growth promoting method of the present invention, the frequency of the AC electric field is 1 uHz or less.

The present invention is a container made of an insulator; Two electrodes attached to the bottom of the container and the outer surface of the lid; And an electric field applying device connected to the two electrodes, wherein the electric field applying device provides an yeast growth promoting device that applies an AC electric field or a DC electric field of 1 Hz or less at an intensity of 1 V to 100 MV / m.

In the yeast growth promoting apparatus of the present invention, the electric field applying device is characterized by applying an alternating electric field of 1uHz or less at a strength of 1V ~ 100MV / m.

In the yeast growth promoting apparatus of the present invention, one of the two electrodes is grounded and the other is characterized in that connected to the anode or cathode of the electric field application device.

In addition, the present invention is a method for producing bioethanol by using the yeast growth promoting method of the present invention, by putting the raw material grains in a container made of an insulator and crushed, pre-treatment by adding a diluted sulfuric acid or amylase enzyme to the grains ; Adding yeast to the pretreated grain; Placing a container containing a mixture of the pretreated grain and yeast between two electrodes; Multiplying yeast by applying an electric field between the two electrodes; And it provides a bio-ethanol manufacturing method comprising the step of distilling the ethanol produced by the yeast growth.

The present invention provides a method for producing an alcoholic beverage using the yeast growth promoting method of the present invention, comprising the steps of glycosylated with maltose or glucose by putting the raw material grains in a container made of an insulator; Adding yeast to the saccharified grains; Placing a container containing the mixed solution of saccharified grains and yeast between two electrodes; Multiplying yeast by applying an electric field between the two electrodes; And after the production of ethanol by the growth of the yeast provides an alcoholic beverage production method comprising the step of removing and aging yeast.

The present invention relates to a method of aging the bread using the yeast growth promoting method of the present invention, comprising the steps of: kneading the flour as a raw material in a container made of an insulator; Adding yeast to the dough; Placing a container containing the yeast-added dough between two electrodes; Multiplying yeast by applying an electric field between the two electrodes; And baking the dough after carbon dioxide is generated by the yeast growth.

It is possible to increase the production of yeast itself by promoting the growth of yeast using the electric field application method and apparatus of the present invention, and to increase the production of metabolites of yeast (eg, ethanol, carbon dioxide). Increasing the production of yeast itself may increase the production of yeast extracts, nutrients, probiotics or other proteins, DNA, etc. present in yeast, and increasing production of yeast metabolites may lead to baking, alcoholic beverage production or bioethanol production. Can increase production efficiency in the industry. In addition, by applying the yeast growth promoting device of the present invention can be devised a yeast culture facility system, aging device, alcohol fermentation system.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail.

The method for promoting yeast growth according to the present invention is a method of applying an electric field to the yeast by a non-contact electric field using a low frequency band electric field of 1 Hz or less, it is applied to all the yeast used industrially to promote growth of 300% or more, Not only does it increase the number of yeasts, but it is also applicable to the production of all metabolites produced in yeast.

The principle that the growth of the yeast is promoted by applying the electric field according to the present invention is as follows. All cells, including yeast, need energy to synthesize the components necessary for cells to grow and divide, and to replicate DNA. This energy is produced and stored in cells in the form of ATP, and if it is not a certain amount, life activity becomes difficult. There are two ways of synthesizing ATP in cells. One method is to break down nutrients such as glucose and convert it into ATP through the respiratory process. The other accumulates hydrogen ions (H +), or protons, outside the cell membrane. When a certain level of proton gradient occurs inside and outside the cell membrane, ATP is generated from ADP while passing protons through ATPase (ATPase) present in the cell membrane. Therefore, the passage of charged particles such as protons by the electric field is promoted, and as a result, the production of ATP increases the rate of cell growth as long as the nutrients necessary for cell growth are present.

The present invention relates to a method for promoting yeast growth by applying a low frequency band electric field of 1 Hz or less to yeast. As an electrode, a metal electrode, a transparent electrode, or an electrode made of a plastic conductive material can be used in various ways. The electrode is connected to an electric field applying device so that one side is grounded and the other is energized. It is done. An AC or DC electric field with a frequency of 1 Hz or less is applied, and the strength of the electric field is not limited, but preferably 1 V to 100 MV / m electric field is used. When applying an alternating electric field, the waveform is not limited, but preferably a sine or square wave is used.

When the electric field is applied to the yeast according to the present invention is characterized in that the electric field is applied to the solution containing the yeast or yeast without contacting the electrode. By applying an electric field in a non-contact manner, the method for promoting yeast growth according to the present invention can be applied to various types of food or yeast liquid mixture. 1 is a schematic diagram of one embodiment of a yeast growth promoting apparatus for applying an electric field in a non-contact manner according to the present invention. In order to apply an electric field in a non-contact manner, a container made of an insulator contains a solution containing yeast or yeast, and the container is placed between two electrodes, and then the electrode is connected to an electric field applying device. And a non-contact electric field application can be carried out by using a container designed to attach two electrodes 2 each made of a conductor material to the outer surface of the lid and connecting the electrodes to the electric field applying device 3.

Hereinafter, the configuration and effects of the present invention will be described in more detail with reference to Examples. However, the following examples are illustrated to aid the understanding of the present invention, and the present invention is not limited only to these examples.

EXAMPLE

First, yeast was mixed in a yeast culture medium and cultured at 1000, 2000, and 10000 CFU / ml. The mixture of the yeast and the culture solution is placed in a 6-well plate made of polypropylene as an insulator, and the sine of 1 kHz, 1 Hz, 1 mHz, and 1 uHz frequency is prevented from directly contacting an electrode applying an electric field as shown in FIG. A wave alternating electric field was applied. The degree of proliferation was evaluated by Optical Density at 600 nm of the mixed solution of the yeast and the culture solution.

According to Figure 2 showing the evaluation results, the yeast growth was promoted when the electric field is applied at a frequency of 1 Hz or less, especially when the electric field is applied when applying the electric field at 1uHz (control group) compared to the growth (3) It was confirmed that ~ 7 times more accelerated. The growth promoting effect may vary depending on the measurement temperature and time.

Useful yeasts that can be used in the present invention include Saccharomyces, Candida, Crebrothecium, Geotrichum, Hansenula, Kloeckera , Lipomyces, Pichia, Rhodosporidium, Rhodotorula, Saccharomycodes, Schizosaccharomyces, Spirobolomyces (Schizosaccharomyces) Yeasts of the genus Sporobolomyces, Torulopsis, Trichosporon and Wickerhamia can be used. As a species included in the above genus, Saccharomyces cerevisiae, Saccharomyces carlsbergensis, Saccharomyces chevalieri, Saccharomyces del Breueki (Saccharomyces delbrueckii), Saccharomyces exiguous, Saccharomyces fermentati, Saccharomyces logos, Saccharomyces mellis, Saccharomyces oviformis, Saccharomyces rosei, Saccharomyces rouxii, Saccharomyces sake, Saccharomyces ubarum Saccharomyces uvarum, Saccharomyces willianus, Saccharomyces sp., Schicharomyces octosporus (Schizosaccha) romyces octosporus, Schizosaccharomyces pombe, Sporobolomyces roseus, Torulopsis candida, Torulopsis famta, Torulopsis globosa (Torulopsis globosa), Torulopsis inconspicua, Trichosporon behrendii, Trichosporon capitatum, Trichosporon cutanum, Wikeramia Wickerhamia fluoresens, Candida arborea, Candida krusei, Candida lambica, Candida lipolytica, Candida parapsilosis, Candida parapsilosis Candida pulcherrima, Candida rugousa, Candida tropicalis, Candida utilis, crebrotesium ash Crebrothecium ashbyii, Geotrichum candidum, Hansenula anomala, Hansenula arabitolgens, Hansenula jadinii, Hansenula satuula satunu ), Hansenula schneggii, Hansenula subpelliculosa, Kloeckera apiculata, Lipomyces starkeyi, Pichia farinose ), Pichia membranaefaciens, Rhodosporidium toruloides, Rhodotorula glutinis, Rhodotorula minuta, Rhodotorula luvar rubar, Rhodotorula aurantiaca, Saccharomycodes ludwigii, and Saccharomycodes si nenses). Particularly useful yeast strains include Saccharomyces cerevisiae Hansen, CGMCC accession No. AS2.375, AS2.501, AS2.502, AS2.503, AS2.504, AS2.535, AS2 .558, AS2.560, AS2.561, AS2.562, or IFF11048); Or Saccharomyces carlsbergensis Hansen, CGMCC accession No. AS2.420 or AS2.444.

1 is a schematic diagram of one embodiment of a yeast growth promoting apparatus according to the present invention.

Figure 2 is a graph showing the results of evaluating the degree of yeast growth by Optical Density after applying the electric field of 1kHz, 1Hz, 1mHz, and 1uHz frequency.

* Description of the symbols for the main parts of the drawings *

1: container 2: electrode

3: electric field applying device

Claims (5)

A yeast growth promoting method comprising applying an electric field to the yeast alternating current or direct current electric field of less than 1 Hz in 1V ~ 100MV / m intensity. The method of claim 1, The frequency of the alternating current electric field is 1uHz or less, characterized in that the yeast growth promoting method. A container 1 made of an insulator; Two electrodes (2) attached to the bottom of the container and the outer surface of the lid; And An electric field applying device 3 connected to the two electrodes, The electric field applying device is a yeast growth promoting device, characterized in that for applying an AC electric field or a DC electric field of less than 1 Hz in 1V ~ 100MV / m intensity. The method of claim 3, The electric field applying device is a yeast growth promoting device, characterized in that for applying an alternating electric field of 1uHz or less in 1V ~ 100MV / m intensity. 4. The yeast growth promoting apparatus according to claim 3, wherein one of the two electrodes is grounded and the other is connected to the anode or the cathode of the electric field applying device.

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